Browsing by Author "Tshoko, Siphokazi"
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Item Green method synthesised graphene-silver electrochemical nanobiosensors for ethambutol and pyrazinamide(MDPI, 2020) Ajay, Rachel. Fanelwa. I.; Tshoko, Siphokazi; Mgwili, YonelaA novel nanobiosensor was constructed with graphene oxide (GO) sheets coupled to pear extract-based green-synthesised silver nanoparticles (Ag-NPs) to which cytochrome P450-2D6 (CYP2D6) enzyme was attached. The biosensor was applied in the electrochemical detection of the tuberculosis (TB) treatment drugs, ethambutol (EMB) and pyrazinamide (PZA). The surface morphology of the green-synthesised nanocomposites was studied by performing High-Resolution Transmission Electron Microscopy (HR-TEM) and High-Resolution Scanning Electron Microscopy (HR-SEM). Fourier Transform Infrared Spectroscopy (FTIR) and Raman Spectroscopy were used for structural analysis, while Ultraviolet Visible (UV-Vis) Spectroscopy was used in the optical characterisation of the nanocomposite material. Electrochemical studies on glassy carbon electrode (GCE), which were done by Cyclic Voltammetry (CV), showed that the GO|Ag-NPs||GCE electrode was highly conductive, and thereby indicating its suitability as a platform for nanobiosensor development. The non-toxic and low-cost green GO|Ag-NPs|CYP2D6||GCE nanobiosensor was used to determine EMB and PZA. The very low limit of detection (LOD) values of the biosensor for EMB (0.2962 × 10−2 nM, S/N = 3) and PZA (0.897 × 10−2 nM, S/N = 3) demonstrate that the green nanobiosensor is more sensitive than other biosensors reported for EMB and PZA.Item Spectroelectrochemical graphene-silver/zinc oxide nanoparticulate phenotype biosensors for ethambutol and pyrazinamide(University of the Western Cape, 2019) Tshoko, Siphokazi; Ajayi, Fanelwa; Iwuoha, Emmanuel; Mulaudzi-Masuku, TakalaniTuberculosis (TB), a deadly disease second to HIV/AIDS, is a global health problem. Diagnosis of active tuberculosis is tedious and requires expensive procedures since there is no recognizable method for sole detection of active TB. Although this is a deadly disease, treatment drug toxicity is also an issue that also causes fatalities in diagnosed patients. Therefore, a rapid sensitive and specific diagnostic method is imperative for TB drug management. In this study spectroscopic and/or electrochemical biosensors were developed for the detection and quantification of TB treatment drugs. The biosensors were constructed with electroactive layers of graphene oxide coupled to silver nanoparticles and/or zinc oxide nanoparticles. These nanoparticles coupled with graphene oxide sheets were covalently attached onto the enzymes such as Cytochrome P450-2D6 to achieve the electrochemical detection of the TB treatment drugs and obtain the required electron transfer between the electrode surface and enzyme. The surface morphology of graphene oxide, nanoparticles as well as the green synthesized nanocomposites were achieved using High-Resolution Transmission Electron Microscopy (HRTEM), Atomic Force Microscopy (AFM), and High- Resolution Scanning Electron Microscopy (HRSEM) while the elemental analysis were obtained using Fourier Transform Infrared Spectroscopy (FTIR), Energy Dispersive X-Ray (EDX), Raman spectroscopy and X-Ray diffraction (XRD). Additionally, the optical properties of the developed nanocomposites where further characterised using Small Angle X-ray Scattering (SAXS), Photoluminescence Spectroscopy (PL) and Ultraviolet Spectroscopy (UV-vis). The electrochemical studies were obtained using cyclic voltammetry (CV) and showed an increase in electron conductivity for the green synthesized zinc oxide nanoparticles coupled with graphene oxide (ZnONPs/GO) and silver nanoparticles coupled with graphene oxide (AgNPs/GO) nanocomposite which was an indication that they were suitable as platforms towards biosensor development. Furthermore, amperometric technique was also used for biotransformation of the TB treatment drugs (Ethambutol and Pyrazinamide) in standard solutions of 0.1 M phosphate buffer (pH 7.0). Furthermore, the sensitivity value of 0.0748 μA/μM was determined for the ethambutol biosensor while a value of 0.1715 μA/μM was determined for the pyrazinamide biosensors. Very good detection limits were obtained for the standard solutions of ethambutol and pyrazinamide where a value of 0.02057 nM was determined for ethambutol at concentration linear range of 50 μM – 400 μM. Additionally, a value of 0.8975 x 10-2 nM was determined for pyrazinamide at the concentration linear range of 100 μM – 300 μM. The determined limit of detections have provided a clear indication that these biosensors have potential of being used in human samples since these values are below the peak serum concentrations of these drugs in TB diagnosed patients as reported in literature. This was further confirmed by the limit of quantification values determined for each biosensor where a value of 0.8975 x 10-2 nM was determined for pyrazinamide and a value of 0.02057 nM was determined for ethambutol.